Sweet corn line SHY-6RSNL196

ABSTRACT

The invention provides seed and plants of corn line SHY-6RSNL196. The invention thus relates to the plants, seeds, and tissue cultures of corn line SHY-6RSNL196 and to methods for producing a corn plant produced by crossing such plants with themselves or with another corn plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the fruit and gametes of such plants.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of inbred corn line SHY-6RSNL196.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial or desirable by a grower and/or consumer, includinggreater yield, resistance to insects or disease, tolerance toenvironmental stress, and nutritional value.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all genetic loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for manygenetic loci. Conversely, a cross of two plants each heterozygous at anumber of loci produces a population of hybrid plants that differgenetically and are not uniform. The resulting non-uniformity makesperformance unpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a corn plant of the cornline designated SHY-6RSNL196. Also provided are corn plants having allthe physiological and morphological characteristics of such a plant.Parts of these corn plants are also provided, for example, includingpollen, an ovule, an embryo, a seed, a scion, a rootstock, a fruit, anda cell of the plant.

In another aspect of the invention, a plant of corn line SHY-6RSNL196comprising an added heritable trait is provided. The heritable trait maycomprise a genetic locus that is, for example, a dominant or recessiveallele. In one embodiment of the invention, a plant of corn lineSHY-6RSNL196 is defined as comprising a single locus conversion. Inspecific embodiments of the invention, an added genetic locus confersone or more traits such as, for example, herbicide tolerance, insectresistance, disease resistance, and modified carbohydrate metabolism. Infurther embodiments, the trait may be conferred by a naturally occurringgene introduced into the genome of a line by backcrossing, a natural orinduced mutation, or a transgene introduced through genetictransformation techniques into the plant or a progenitor of any previousgeneration thereof. When introduced through transformation, a geneticlocus may comprise one or more genes integrated at a single chromosomallocation.

In some embodiments, a single locus conversion includes one or moresite-specific changes to the plant genome, such as, without limitation,one or more nucleotide modifications, deletions, or insertions. A singlelocus may comprise one or more genes or nucleotides integrated ormutated at a single chromosomal location. In one embodiment, a singlelocus conversion may be introduced by a genetic engineering technique,methods of which include, for example, genome editing with engineerednucleases (GEEN). Engineered nucleases include, but are not limited to,Cas endonucleases; zinc finger nucleases (ZFNs); transcriptionactivator-like effector nucleases (TALENs); engineered meganucleases,also known as homing endonucleases; and other endonucleases for DNA orRNA-guided genome editing that are well-known to the skilled artisan.

The invention also concerns the seed of corn line SHY-6RSNL196. The cornseed of the invention may be provided as an essentially homogeneouspopulation of corn seed of corn line SHY-6RSNL196. Essentiallyhomogeneous populations of seed are generally free from substantialnumbers of other seed. Therefore, seed of corn line SHY-6RSNL196 may bedefined as forming at least about 97% of the total seed, including atleast about 98%, 99% or more of the seed. The seed population may beseparately grown to provide an essentially homogeneous population ofcorn plants designated SHY-6RSNL196.

In yet another aspect of the invention, a tissue culture of regenerablecells of a corn plant of corn line SHY-6RSNL196 is provided. The tissueculture will preferably be capable of regenerating corn plants capableof expressing all of the physiological and morphological characteristicsof the starting plant and of regenerating plants having substantiallythe same genotype as the starting plant. Examples of some of thephysiological and morphological characteristics of corn lineSHY-6RSNL196 include those traits set forth in the tables herein. Theregenerable cells in such tissue cultures may be derived, for example,from embryos, meristems, cotyledons, pollen, leaves, anthers, roots,root tips, pistils, flowers, seeds, stalks, silks, tassels, kernels,ears, cobs, and husks. Still further, the present invention providescorn plants regenerated from a tissue culture of the invention, theplants having all the physiological and morphological characteristics ofcorn line SHY-6RSNL196.

In still yet another aspect of the invention, processes are provided forproducing corn seeds, plants, and fruit, which processes generallycomprise crossing a first parent corn plant with a second parent cornplant, wherein at least one of the first or second parent corn plants isa plant of corn line SHY-6RSNL196. These processes may be furtherexemplified as processes for preparing hybrid corn seed or plants,wherein a first corn plant is crossed with a second corn plant of adifferent, distinct genotype to provide a hybrid that has, as one of itsparents, a plant of corn line SHY-6RSNL196. In these processes, crossingwill result in the production of seed. The seed production occursregardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent corn plant, oftenin proximity so that pollination will occur for example, mediated byinsect vectors. Alternatively, pollen can be transferred manually. Wherethe plant is self-pollinated, pollination may occur without the need fordirect human intervention other than plant cultivation. For hybridcrosses, it may be beneficial to detassel or otherwise emasculate theparent used as a female.

A second step may comprise cultivating or growing the seeds of first andsecond parent corn plants into mature plants. A third step may comprisepreventing self-pollination of the plants, such as by detasseling orother means.

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent corn plants. Yet another step comprisesharvesting the seeds from at least one of the parent corn plants. Theharvested seed can be grown to produce a corn plant or hybrid cornplant.

The present invention also provides the corn seeds and plants producedby a process that comprises crossing a first parent corn plant with asecond parent corn plant, wherein at least one of the first or secondparent corn plants is a plant of corn line SHY-6RSNL196. In oneembodiment of the invention, corn seed and plants produced by theprocess are first generation (F₁) hybrid corn seed and plants producedby crossing a plant in accordance with the invention with another,distinct plant. The present invention further contemplates plant partsof such an F₁ hybrid corn plant and methods of use thereof. Therefore,certain exemplary embodiments of the invention provide an F₁ hybrid cornplant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from corn line SHY-6RSNL196, the methodcomprising the steps of: (a) preparing a progeny plant derived from cornline SHY-6RSNL196, wherein said preparing comprises crossing a plant ofcorn line SHY-6RSNL196 with a second plant; and (b) crossing the progenyplant with itself or a second plant to produce a seed of a progeny plantof a subsequent generation. In further embodiments, the method mayadditionally comprise: (c) growing a progeny plant of a subsequentgeneration from said seed of a progeny plant of a subsequent generationand crossing the progeny plant of a subsequent generation with itself ora second plant; and repeating the steps for an additional 3-10generations to produce a plant derived from corn line SHY-6RSNL196. Theplant derived from corn line SHY-6RSNL196 may be an inbred line, and theaforementioned repeated crossing steps may be defined as comprisingsufficient inbreeding to produce the inbred line. In the method, it maybe desirable to select particular plants resulting from step (c) forcontinued crossing according to steps (b) and (c). By selecting plantshaving one or more desirable traits, a plant derived from corn lineSHY-6RSNL196 is obtained which possesses some of the desirable traits ofthe line/hybrid as well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of corn lineSHY-6RSNL196, wherein the plant has been cultivated to maturity, and (b)collecting at least one corn from the plant.

In still yet another aspect of the invention, the genetic complement ofcorn line SHY-6RSNL196 is provided. The phrase “genetic complement” isused to refer to the aggregate of nucleotide sequences, the expressionof which sequences defines the phenotype of, in the present case, a cornplant, or a cell or tissue of that plant. A genetic complement thusrepresents the genetic makeup of a cell, tissue, or plant, and a hybridgenetic complement represents the genetic make-up of a hybrid cell,tissue, or plant. The invention thus provides corn plant cells that havea genetic complement in accordance with the corn plant cells disclosedherein and seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles andby the expression of phenotypic traits that are characteristic of theexpression of the genetic complement, e.g., isozyme typing profiles. Itis understood that corn line SHY-6RSNL196 could be identified by any ofthe many well-known techniques such as, for example, Simple SequenceLength Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 18:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR),Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858,specifically incorporated herein by reference in its entirety), andSingle Nucleotide Polymorphisms (SNPs) (Wang et al., Science,280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by corn plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a corn plant of the invention with a haploid geneticcomplement of a second corn plant, preferably, another, distinct cornplant. In another aspect, the present invention provides a corn plantregenerated from a tissue culture that comprises a hybrid geneticcomplement of this invention.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features, and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds, and derivatives of corn line SHY-6RSNL196. This line showsuniformity and stability within the limits of environmental influencefor the traits described hereinafter. Corn line SHY-6RSNL196 providessufficient seed yield. By crossing with a distinct second plant, uniformF₁ progeny can be obtained.

A. Physiological and Morphological Characteristics of Corn Line SHY-6RSNL196

In accordance with one aspect of the present invention, there areprovided plants having the physiological and morphologicalcharacteristics of corn line SHY-6RSNL196. A description of thephysiological and morphological characteristics of such plants ispresented in Table 1.

TABLE 1 Physiological and Morphological Characteristics of Corn LineSHY-6RSNL196 CHARACTERISTIC SHY-6RSNL196 Florida 32 Type sweet sweetRegion Where Developed in the northwest northwest U.S.A. Maturity in theRegion of Best Adaptability emergence to 50% of plants in silk    58days   49 days  946.5 heat units  1103 heat units emergence to 50% ofplants in pollen    55 days   47 days   889 heat units  1053 heat units10% to 90% pollen shed    9 days   18 days  226.9 heat units 438.4 heatunits 50% silk to optimum edible quality    16 days   15 days 384.05heat units 362.4 heat units Plant plant height (to tassel tip) (cm) avg:141.84 avg: 135.27 std dev: 11.8785 std dev: 17.7333 sample size: 15sample size: 15 ear height (to base of top ear node) avg: 45.17 cm avg:30.23 (cm) std dev: 2.9196 std dev: 7.3092 sample size: 15 sample size:15 length of top ear internode (cm) avg: 13.05 avg: 10.17 std dev:0.8782 std dev: 1.3151 sample size: 15 sample size: 15 average number oftillers avg: 1.07 avg: 3.07 std dev: 0.8837 std dev: 1.6242 sample size:15 sample size: 15 average number of ears per stalk avg: 1.93 avg: 2.73std dev: 0.5936 std dev: 1.1629 sample size: 15 sample size: 15anthocyanin of brace roots absent absent Leaf width of ear node leaf(cm) avg: 5.85 avg: 6.68 std dev: 0.4734 std dev: 0.9104 sample size: 15sample size: 15 length of ear node leaf (cm) avg: 61.15 avg: 72.89 stddev: 2.85 std dev: 4.103 sample size: 15 sample size: 15 number ofleaves above top ear avg: 4.33 avg: 4.07 std dev: 0.488 std dev: 0.2582sample size: 15 sample size: 15 leaf angle (second leaf above ear at70.5 50 anthesis to stalk above leaf) (degrees) color (Munsell ColorChart Code) 5GY 4/6 5GY 4/6 leaf sheath pubescence (scale from 1  6  2(none) to 9 (like peach fuzz)) marginal waves (scale from 1 (none)  7  5to 9 (many)) longitudinal creases (scale from 1  8  8 (none) to 9(many)) Tassel number of primary lateral branches avg: 11.67 avg: 12.33std dev: 1.7182 std dev: 3.2878 sample size: 15 sample size: 15 branchangle from central spike avg: 66.67 avg: 52.67 (degrees) std dev: 8.5161std dev: 9.0053 sample size: 15 sample size: 15 tassel length (top leafcollar to tassel avg: 30.13 avg: 45.05 tip) (cm) std dev: 2.941 std dev:3.8726 sample size: 15 sample size: 15 pollen shed (scale from 0 (male 7  8 sterile) to 9 (heavy shed)) anther color (Munsell Color Chart2.5GY 8/6 2.5GY 8/6 Code) glume color (Munsell Color Chart   5GY 6/4  5GY 5/4 Code) bar glumes (glume bands) present present Ear silk color(unhusked) (Munsell Color 2.5 8/6 2.5GY 8/6 Chart Code) (3 days afteremergence) fresh husk color (unhusked) (Munsell 2.5GY 7/4   5GY 7/6Color Chart Code) (25 days after 50% silking) husk tightness (unhusked)(scale from  4  6 1 (very loose) to 9 (very tight)) husk extension(unhusked) (at harvest) long medium ear length (husked) (cm) avg: 14.99avg: 16.19 std dev: 1.0586 std dev: 2.5144 sample size: 15 sample size:15 ear diameter at mid-point (husked) avg: 32.01 avg: 35.71 (mm) stddev: 2.6451 std dev: 2.4195 sample size: 15 sample size: 15 ear weight(husked) (g) avg: 62.67 avg: 78.73 std dev: 14.7487 std dev: 30.6954sample size: 15 sample size: 15 number of kernel rows (husked) avg: 15.2avg: 17.33 std dev: 1.5213 std dev: 1.3973 sample size: 15 sample size:15 kernel rows (husked) indistinct indistinct row alignment (husked)slightly curved slightly curved shank length (husked) (cm) avg: 11.15avg: 27.21 std dev: 1.9464 std dev: 3.5522 sample size: 15 sample size:15 ear taper (husked) average average Cob cob diameter at mid-point (mm)avg: 25.9 avg: 27.33 std dev: 1.59 std dev: 1.8836 sample size: 15sample size: 15 cob color (Munsell Color Chart Code) 2.5Y 8/2 2.5Y 8/4Agronomic Traits pre-anthesis brittle snapping  0%  0% pre-anthesis rootlodging  0%  0% These are typical values. Values may vary due toenvironment. Other values that are substantially equivalent are withinthe scope of the invention.These are typical values. Values may vary due to environment. Othervalues that are substantially equivalent are within the scope of theinvention.

B. Breeding Corn Plants

In one aspect of the current invention line SHY-6RSNL196 may be crossedwith itself or with any second plant. Such methods can be used forpropagation of the corn line SHY-6RSNL196 or can be used to produceplants that are derived from the corn line SHY-6RSNL196. Plants derivedfrom the corn line SHY-6RSNL196 may be used, in certain embodiments, forthe development of new corn varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing a plant of the invention followed by multiplegenerations of breeding according to such well-known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid induction systems have been developedfor various plants to produce haploid tissues, plants and seeds. Thehaploid induction system can produce haploid plants from any genotype bycrossing with an inducer line. Inducer lines and methods for obtaininghaploid plants are known in the art.

Haploid embryos may be produced, for example, from microspores, pollen,anther cultures, or ovary cultures. The haploid embryos may then bedoubled autonomously, or by chemical treatments (e.g. colchicinetreatment). Alternatively, haploid embryos may be grown into haploidplants and treated to induce chromosome doubling. In either case,fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with a plantof the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the plants. In selecting a second plant to cross with cornline SHY-6RSNL196 for the purpose of developing novel corn lines, itwill typically be preferred to choose those plants which eitherthemselves exhibit one or more selected desirable characteristics orwhich exhibit the desired characteristic(s) when in hybrid combination.Examples of desirable traits may include, in specific embodiments, malesterility, herbicide resistance, resistance for bacterial, fungal, orviral disease, insect resistance, male fertility, sugar content, andenhanced nutritional quality.

C. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing or by genetic engineering, whereinessentially all of the morphological and physiological characteristicsof a variety are recovered in addition to a genetic locus transferredinto the plant via the backcrossing or genetic engineering technique.The term single locus converted plant as used herein refers to thosecorn plants which are developed by a plant breeding technique calledbackcrossing or by genetic engineering, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered or conserved in addition to the single locus introduced intothe variety via the backcrossing or genetic engineering technique,respectively. By essentially all of the morphological and physiologicalcharacteristics, it is meant that the characteristics of a plant arerecovered or conserved that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing, introduction of a transgene, or applicationof a genetic engineering technique.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalcorn plant which contributes the locus for the desired characteristic istermed the nonrecurrent or donor parent. This terminology refers to thefact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental corn plant to whichthe locus or loci from the nonrecurrent parent are transferred is knownas the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a corn plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny corn plants of a backcross in which a plantdescribed herein is the recurrent parent comprise (i) the desired traitfrom the non-recurrent parent and (ii) all of the physiological andmorphological characteristics of corn the recurrent parent as determinedat the 5% significance level when grown in the same environmentalconditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiol., 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into an eliteline. This molecular breeding-facilitated movement of a trait or traitsinto an elite line may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the eliteline by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into an elite line via this methodology. Whenthis elite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,male sterility, waxy starch, herbicide resistance, resistance tobacterial, fungal, or viral disease, insect resistance, sugar content,male fertility and enhanced nutritional quality. These genes aregenerally inherited through the nucleus, but may be inherited throughthe cytoplasm. Some known exceptions to this are genes for malesterility, some of which are inherited cytoplasmically, but still act asa single locus trait.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of corn plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. In addition,marker assisted selection may be used to identify plants comprisingdesirable genotypes at the seed, seedling, or plant stage, to identifyor assess the purity of a cultivar, to catalog the genetic diversity ofa germplasm collection, and to monitor specific alleles or haplotypeswithin an established cultivar.

Types of genetic markers which could be used in accordance with theinvention include, but are not necessarily limited to, Simple SequenceLength Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 18:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR),Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858,specifically incorporated herein by reference in its entirety), andSingle Nucleotide Polymorphisms (SNPs) (Wang et al., Science,280:1077-1082, 1998).

In particular embodiments of the invention, marker assisted selection isused to increase the efficiency of a backcrossing breeding scheme forproducing a corn line comprising a desired trait. This technique iscommonly referred to as marker assisted backcrossing (MABC). Thistechnique is well-known in the art and may involve, for example, the useof three or more levels of selection, including foreground selection toidentity the presence of a desired locus, which may complement orreplace phenotype screening protocols; recombinant selection to minimizelinkage drag; and background selection to maximize recurrent parentgenome recovery.

D. Plants Derived by Genetic Engineering

Various genetic engineering technologies have been developed and may beused by those of skill in the art to introduce traits in plants. Incertain aspects of the claimed invention, traits are introduced intocorn plants via altering or introducing a single genetic locus ortransgene into the genome of a recited variety or progenitor thereof.Methods of genetic engineering to modify, delete, or insert genes andpolynucleotides into the genomic DNA of plants are well-known in theart.

In specific embodiments of the invention, improved corn lines can becreated through the site-specific modification of a plant genome.Methods of genetic engineering include, for example, utilizingsequence-specific nucleases such as zinc-finger nucleases (see, forexample, U.S. Pat. Appl. Pub. No. 2011-0203012); engineered or nativemeganucleases; TALE-endonucleases (see, for example, U.S. Pat. Nos.8,586,363 and 9,181,535); and RNA-guided endonucleases, such as those ofthe CRISPR/Cas systems (see, for example, U.S. Pat. Nos. 8,697,359 and8,771,945 and U.S. Pat. Appl. Pub. No. 2014-0068797). One embodiment ofthe invention thus relates to utilizing a nuclease or any associatedprotein to carry out genome modification. This nuclease could beprovided heterologously within donor template DNA for templated-genomicediting or in a separate molecule or vector. A recombinant DNA constructmay also comprise a sequence encoding one or more guide RNAs to directthe nuclease to the site within the plant genome to be modified. Furthermethods for altering or introducing a single genetic locus include, forexample, utilizing single-stranded oligonucleotides to introduce basepair modifications in a corn plant genome (see, for example Sauer etal., Plant Physiol, 170(4):1917-1928, 2016).

Methods for site-directed alteration or introduction of a single geneticlocus are well-known in the art and include those that utilizesequence-specific nucleases, such as the aforementioned, or complexes ofproteins and guide-RNA that cut genomic DNA to produce a double-strandbreak (DSB) or nick at a genetic locus. As is well-understood in theart, during the process of repairing the DSB or nick introduced by thenuclease enzyme, a donor template, transgene, or expression cassettepolynucleotide may become integrated into the genome at the site of theDSB or nick. The presence of homology arms in the DNA to be integratedmay promote the adoption and targeting of the insertion sequence intothe plant genome during the repair process through homologousrecombination or non-homologous end joining (NHEJ).

In another embodiment of the invention, genetic transformation may beused to insert a selected transgene into a plant of the invention ormay, alternatively, be used for the preparation of transgenes which canbe introduced by backcrossing. Methods for the transformation of plantsthat are well-known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation, anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and, preferably, gold. For the bombardment, cells insuspension are concentrated on filters or solid culture medium.Alternatively, immature embryos or other target cells may be arranged onsolid culture medium. The cells to be bombarded are positioned at anappropriate distance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Nat. Biotechnol., 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Nat. Biotechnol., 3:629-635, 1985; U.S.Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994) and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985),including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591,1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S); the nopaline synthase promoter (An et al., Plant Physiol.,88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell,1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem;the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform viruspromoter; a commelina yellow mottle virus promoter; and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., PlantPhysiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter,Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-bindingprotein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones,such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4)wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a corn plant according to the invention.Non-limiting examples of particular genes and corresponding phenotypesone may choose to introduce into a corn plant include one or more genesfor insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pesttolerance such as genes for fungal disease control, herbicide tolerancesuch as genes conferring glyphosate tolerance and genes for qualityimprovements such as yield, nutritional enhancements, environmental orstress tolerances, or any desirable changes in plant physiology, growth,development, morphology or plant product(s). For example, structuralgenes would include any gene that confers insect tolerance including butnot limited to a Bacillus insect control protein gene as described in WO99/31248, herein incorporated by reference in its entirety, U.S. Pat.No. 5,689,052, herein incorporated by reference in its entirety, U.S.Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference intheir entirety. In another embodiment, the structural gene can confertolerance to the herbicide glyphosate as conferred by genes including,but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPSgene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, hereinincorporated by reference in its entirety, or glyphosate oxidoreductasegene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporatedby reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

E. Male Sterility

Examples of genes conferring male sterility include those disclosed inU.S. Pat. Nos. 3,861,709, 3,710,511, 4,654,465, 5,625,132, and4,727,219, each of the disclosures of which are specificallyincorporated herein by reference in their entirety. Male sterility genescan increase the efficiency with which hybrids are made, in that theyeliminate the need to physically emasculate the corn plant used as afemale in a given cross.

Where one desires to employ male-sterility systems with a corn plant inaccordance with the invention, it may be beneficial to also utilize oneor more male-fertility restorer genes. For example, where cytoplasmicmale sterility (CMS) is used, hybrid seed production requires threeinbred lines: (1) a cytoplasmically male-sterile line having a CMScytoplasm; (2) a fertile inbred with normal cytoplasm, which is isogenicwith the CMS line for nuclear genes (“maintainer line”); and (3) adistinct, fertile inbred with normal cytoplasm, carrying a fertilityrestoring gene (“restorer” line). The CMS line is propagated bypollination with the maintainer line, with all of the progeny being malesterile, as the CMS cytoplasm is derived from the female parent. Thesemale sterile plants can then be efficiently employed as the femaleparent in hybrid crosses with the restorer line, without the need forphysical emasculation of the male reproductive parts of the femaleparent.

The presence of a male-fertility restorer gene results in the productionof fully fertile F₁ hybrid progeny. If no restorer gene is present inthe male parent, male-sterile hybrids are obtained. Such hybrids areuseful where the vegetative tissue of the corn plant is utilized, e.g.,for silage, but in most cases, the seeds will be deemed the mostvaluable portion of the crop, so fertility of the hybrids in these cropsmust be restored. Therefore, one aspect of the current inventionconcerns a corn plant of the invention comprising a genetic locuscapable of restoring male fertility in an otherwise male-sterile plant.Examples of male-sterility genes and corresponding restorers which couldbe employed with the plants of the invention are well known to those ofskill in the art of plant breeding and are disclosed in, for instance,U.S. Pat. Nos. 5,530,191; 5,689,041; 5,741,684; and 5,684,242, thedisclosures of which are each specifically incorporated herein byreference in their entirety.

F. Herbicide Resistance

Numerous herbicide resistance genes are known and may be employed withthe invention. An example is a gene conferring resistance to a herbicidethat inhibits the growing point or meristem, such as an imidazalinone ora sulfonylurea. Exemplary genes in this category code for mutant ALS andAHAS enzyme as described, for example, by Lee et al., EMBO J., 7:1241,1988; Gleen et al., Plant Molec. Biology, 18:1185-1187, 1992; and Mikiet al., Theor. Appl. Genet., 80:449, 1990.

Resistance genes for glyphosate (resistance conferred by mutant5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively), and hygromycin B phosphotransferase, and to otherphosphono compounds such as glufosinate (phosphinothricin acetyltransferase (PAT) and Streptomyces hygroscopicus phosphinothricin-acetyltransferase (bar) genes) may also be used. See, for example, U.S. Pat.No. 4,940,835 to Shah et al., which discloses the nucleotide sequence ofa form of EPSPS which can confer glyphosate resistance. A DNA moleculeencoding a mutant aroA gene can be obtained under ATCC accession number39256, and the nucleotide sequence of the mutant gene is disclosed inU.S. Pat. No. 4,769,061 to Comai. A hygromycin B phosphotransferase genefrom E. coli that confers resistance to glyphosate in tobacco callus andplants is described in Penaloza-Vazquez et al. (Plant Cell Reports,14:482-487, 1995). European patent application No. 0 333 033 to Kumadaet al., and U.S. Pat. No. 4,975,374 to Goodman et al., disclosenucleotide sequences of glutamine synthetase genes which conferresistance to herbicides such as L-phosphinothricin. The nucleotidesequence of a phosphinothricin-acetyltransferase gene is provided inEuropean application No. 0 242 246 to Leemans et al. DeGreef et al.,(Biotechnology, 7:61, 1989), describe the production of transgenicplants that express chimeric bar genes coding for phosphinothricinacetyl transferase activity. Exemplary of genes conferring resistance tophenoxy propionic acids and cyclohexanediones, such as sethoxydim andhaloxyfop are the Acct-S1, Accl-S2 and Acct-S3 genes described byMarshall et al., (Theor. Appl. Genet., 83:4:35, 1992).

Genes conferring resistance to a herbicide that inhibits photosynthesisare also known, such as a triazine (psbA and gs+ genes) and abenzonitrile (nitrilase gene). Przibilla et al., (Plant Cell, 3:169,1991), describe the transformation of Chlamydomonas with plasmidsencoding mutant psbA genes. Nucleotide sequences for nitrilase genes aredisclosed in U.S. Pat. No. 4,810,648 to Stalker, and DNA moleculescontaining these genes are available under ATCC Accession Nos. 53435,67441, and 67442. Cloning and expression of DNA coding for a glutathioneS-transferase is described by Hayes et al., (Biochem. J., 285(Pt1):173-180, 1992). Protoporphyrinogen oxidase (PPO) is the target of thePPO-inhibitor class of herbicides; a PPO-inhibitor resistant PPO genewas recently identified in Amaranthus tuberculatus (Patzoldt et al.,PNAS, 103(33):12329-2334, 2006). The herbicide methyl viologen inhibitsCO₂ assimilation. Foyer et al. (Plant Physiol., 109:1047-1057, 1995)describe a plant overexpressing glutathione reductase (GR) which isresistant to methyl viologen treatment.

Siminszky (Phytochemistry Reviews, 5:445-458, 2006) describes plantcytochrome P450-mediated detoxification of multiple, chemicallyunrelated classes of herbicides.

G. Waxy Starch

The waxy characteristic is an example of a recessive trait. In thisexample, the progeny resulting from the first backcross generation (BC1)must be grown and selfed. A test is then run on the selfed seed from theBC1 plant to determine which BC1 plants carried the recessive gene forthe waxy trait. In other recessive traits additional progeny testing,for example growing additional generations such as the BC1S1, may berequired to determine which plants carry the recessive gene.

H. Disease Resistance

Plant defenses are often activated by specific interaction between theproduct of a disease resistance gene (R) in the plant and the product ofa corresponding avirulence (Avr) gene in the pathogen. A plant line canbe transformed with a cloned resistance gene to engineer plants that areresistant to specific pathogen strains. See, for example, Jones et al.,Science, 266:7891, 1994 (cloning of the tomato Cf-9 gene for resistanceto Cladosporium flavum); Martin et al., Science, 262: 1432, 1993 (tomatoPto gene for resistance to Pseudomonas syringae pv.); and Mindrinos etal., Cell, 78(6):1089-1099, 1994 (Arabidopsis RPS2 gene for resistanceto Pseudomonas syringae).

A viral-invasive protein or a complex toxin derived therefrom may alsobe used for viral disease resistance. For example, the accumulation ofviral coat proteins in transformed plant cells imparts resistance toviral infection and/or disease development effected by the virus fromwhich the coat protein gene is derived, as well as by related viruses.See Beachy et al., (Ann. Rev. Phytopathol., 28:451, 1990). Coatprotein-mediated resistance has been conferred upon transformed plantsagainst alfalfa mosaic virus, cucumber mosaic virus, tobacco streakvirus, potato virus X, potato virus Y, tobacco etch virus, tobaccorattle virus and tobacco mosaic virus. Id.

A virus-specific antibody may also be used. See, for example,Tavladoraki et al., (Nature, 366:469, 1993), who show that transgenicplants expressing recombinant antibody genes are protected from virusattack. Additional means of inducing whole-plant resistance to apathogen include modulation of the systemic acquired resistance (SAR) orpathogenesis related (PR) genes, for example genes homologous to theArabidopsis thaliana NIM1/NPR1/SA11, and/or by increasing salicylic acidproduction (Ryals et al., Plant Cell, 8:1809-1819, 1996).

Logemann et al., (Biotechnology, 10:305, 1992), for example, disclosetransgenic plants expressing a barley ribosome-inactivating gene have anincreased resistance to fungal disease. Plant defensins may be used toprovide resistance to fungal pathogens (Thomma et al., Planta,216:193-202, 2002). Other examples of fungal disease resistance areprovided in U.S. Pat. Nos. 6,653,280; 6,573,361; 6,506,962; 6,316,407;6,215,048; 5,516,671; 5,773,696; 6,121,436; 6,316,407; and 6,506,962.

I. Insect Resistance

One example of an insect resistance gene includes a Bacillusthuringiensis (Bt) protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., (Gene,48:109-118, 1986), who disclose the cloning and nucleotide sequence of aBt δ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genescan be purchased from the American Type Culture Collection, Manassas,Va., for example, under ATCC Accession Nos. 40098, 67136, 31995 and31998. Another example is a lectin. See, for example, Van Damme et al.,(Plant Molec. Biol., 24:25, 1994), who disclose the nucleotide sequencesof several Clivia miniata mannose-binding lectin genes. Avitamin-binding protein may also be used, such as avidin. See PCTapplication US93/06487, the contents of which are hereby incorporated byreference. This application teaches the use of avidin and avidinhomologues as larvicides against insect pests.

Yet another insect resistance gene is an enzyme inhibitor, for example,a protease or proteinase inhibitor or an amylase inhibitor. See, forexample, Abe et al., (J. Biol. Chem., 262:16793, 1987) (nucleotidesequence of rice cysteine proteinase inhibitor), Huub et al., (PlantMolec. Biol., 21:985, 1993) (nucleotide sequence of cDNA encodingtobacco proteinase inhibitor I), and Sumitani et al., (Biosci. Biotech.Biochem., 57:1243, 1993) (nucleotide sequence of Streptomycesnitrosporeus α-amylase inhibitor).

An insect-specific hormone or pheromone may also be used. See, forexample, the disclosure by Hammock et al., (Nature, 344:458, 1990), ofbaculovirus expression of cloned juvenile hormone esterase, aninactivator of juvenile hormone, Gade and Goldsworthy (Eds.Physiological System in Insects, Elsevier Academic Press, Burlington,Mass., 2007), describing allostatins and their potential use in pestcontrol; and Palli et al., (Vitam. Horm., 73:59-100, 2005), disclosinguse of ecdysteroid and ecdysteroid receptor in agriculture. The diuretichormone receptor (DHR) was identified in Price et al., (Insect Mol.Biol., 13:469-480, 2004) as a candidate target of insecticides.

Still other examples include an insect-specific antibody or animmunotoxin derived therefrom and a developmental-arrestive protein. SeeTaylor et al., (Seventh Int'l Symposium on Molecular Plant-MicrobeInteractions, Edinburgh, Scotland, Abstract W97, 1994), who describedenzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments.

Nematode resistance has been described, for example, in U.S. Pat. No.6,228,992 and bacterial disease resistance in U.S. Pat. No. 5,516,671.

J. Modified Fatty Acid, Phytate, and Carbohydrate Metabolism

Genes may be used conferring modified fatty acid metabolism. Forexample, stearyl-ACP desaturase genes may be used. See Knutzon et al.,(Proc. Natl. Acad. Sci. USA, 89:2624, 1992). Various fatty aciddesaturases have also been described, such as a Saccharomyces cerevisiaeOLE1 gene encoding Δ9 fatty acid desaturase, an enzyme which forms themonounsaturated palmitoleic (16:1) and oleic (18:1) fatty acids frompalmitoyl (16:0) or stearoyl (18:0) CoA (McDonough et al., J. Biol.Chem., 267(9):5931-5936, 1992); a gene encoding a stearoyl-acyl carrierprotein delta-9 desaturase from castor (Fox et al., Proc. Natl. Acad.Sci. USA, 90(6):2486-2490, 1993); Δ6- and Δ12-desaturases from thecyanobacteria Synechocystis responsible for the conversion of linoleicacid (18:2) to gamma-linolenic acid (18:3 gamma) (Reddy et al., PlantMol. Biol., 22(2):293-300, 1993); a gene from Arabidopsis thaliana thatencodes an omega-3 desaturase (Arondel et al., Science,258(5086):1353-1355 1992); plant Δ9-desaturases (PCT Application Publ.No. WO 91/13972) and soybean and Brassica Δ15 desaturases (EuropeanPatent Application Publ. No. EP 0616644).

Phytate metabolism may also be modified by introduction of aphytase-encoding gene to enhance breakdown of phytate, adding more freephosphate to the transformed plant. For example, see Van Hartingsveldtet al., (Gene, 127:87, 1993), for a disclosure of the nucleotidesequence of an Aspergillus niger phytase gene. In corn, this, forexample, could be accomplished by cloning and then reintroducing DNAassociated with the single allele which is responsible for corn mutantscharacterized by low levels of phytic acid. See Raboy et al., PlantPhysiol., 124(1):355-368, 1990.

A number of genes are known that may be used to alter carbohydratemetabolism. For example, plants may be transformed with a gene codingfor an enzyme that alters the branching pattern of starch. See Shirozaet al., (J. Bacteriol., 170:810, 1988) (nucleotide sequence ofStreptococcus mutans fructosyltransferase gene), Steinmetz et al., (Mol.Gen. Genet., 20:220, 1985) (nucleotide sequence of Bacillus subtilislevansucrase gene), Pen et al., (Biotechnology, 10:292, 1992)(production of transgenic plants that express Bacillus licheniformisα-amylase), Elliot et al., (Plant Molec. Biol., 21:515, 1993)(nucleotide sequences of tomato invertase genes), Sergaard et al., (J.Biol. Chem., 268:22480, 1993) (site-directed mutagenesis of barleyα-amylase gene), and Fisher et al., (Plant Physiol., 102:1045, 1993)(maize endosperm starch branching enzyme II). The Z10 gene encoding a 10kD zein storage protein from maize may also be used to alter thequantities of 10 kD zein in the cells relative to other components(Kirihara et al., Gene, 71(2):359-370, 1988).

U.S. Pat. No. 6,930,225 describes maize cellulose synthase genes andmethods of use thereof.

K. Resistance to Abiotic Stress

Abiotic stress includes dehydration or other osmotic stress, salinity,high or low light intensity, high or low temperatures, submergence,exposure to heavy metals, and oxidative stress.Delta-pyrroline-5-carboxylate synthetase (P5CS) from mothbean has beenused to provide protection against general osmotic stress.Mannitol-1-phosphate dehydrogenase (mt1D) from E. coli has been used toprovide protection against drought and salinity. Choline oxidase (codAfrom Arthrobactor globiformis) can protect against cold and salt. E.coli choline dehydrogenase (betA) provides protection against salt.Additional protection from cold can be provided by omega-3-fatty aciddesaturase (fad7) from Arabidopsis thaliana. Trehalose-6-phosphatesynthase and levan sucrase (SacB) from yeast and Bacillus subtilis,respectively, can provide protection against drought (summarized fromAnnex II Genetic Engineering for Abiotic Stress Tolerance in Plants,Consultative Group On International Agricultural Research TechnicalAdvisory Committee). Overexpression of superoxide dismutase can be usedto protect against superoxides, as described in U.S. Pat. No. 5,538,878to Thomas et al.

L. Additional Traits

Additional traits can be introduced into a corn variety of the presentinvention. A non-limiting example of such a trait is a coding sequencewhich decreases RNA and/or protein levels. The decreased RNA and/orprotein levels may be achieved through RNAi methods, such as thosedescribed in U.S. Pat. No. 6,506,559 to Fire and Mellow.

Another trait that may find use with the corn variety of the inventionis a sequence which allows for site-specific recombination. Examples ofsuch sequences include the FRT sequence, used with the FLP recombinase(Zhu and Sadowski, J. Biol. Chem., 270:23044-23054, 1995); and the LOXsequence, used with CRE recombinase (Sauer, Mol. Cell. Biol.,7:2087-2096, 1987). The recombinase genes can be encoded at any locationwithin the genome of the corn plant, and are active in the hemizygousstate.

It may also be desirable to make corn plants more tolerant to or moreeasily transformed with Agrobacterium tumefaciens. Expression of p53 andiap, two baculovirus cell-death suppressor genes, inhibited tissuenecrosis and DNA cleavage. Additional targets can include plant-encodedproteins that interact with the Agrobacterium Vir genes; enzymesinvolved in plant cell wall formation; and histones, histoneacetyltransferases and histone deacetylases (reviewed in Gelvin,Microbiology & Mol. Biol. Reviews, 67:16-37, 2003).

In addition to the modification of oil, fatty acid or phytate contentdescribed above, it may additionally be beneficial to modify the amountsor levels of other compounds. For example, the amount or composition ofantioxidants can be altered. See, for example, U.S. Pat. Nos. 6,787,618and 7,154,029 and International Patent Appl. Pub. No. WO 00/68393, whichdisclose the manipulation of antioxidant levels, and InternationalPatent Appl. Pub. No. WO 03/082899, which discloses the manipulation ofan antioxidant biosynthetic pathway.

Additionally, seed amino acid content may be manipulated. U.S. Pat. No.5,850,016 and International Patent Appl. Pub. No. WO 99/40209 disclosethe alteration of the amino acid compositions of seeds. U.S. Pat. Nos.6,080,913 and 6,127,600 disclose methods of increasing accumulation ofessential amino acids in seeds.

U.S. Pat. No. 5,559,223 describes synthetic storage proteins in whichthe levels of essential amino acids can be manipulated. InternationalPatent Appl. Pub. No. WO 99/29882 discloses methods for altering aminoacid content of proteins. International Patent Appl. Pub. No. WO98/20133 describes proteins with enhanced levels of essential aminoacids. International Patent Appl. Pub. No. WO 98/56935 and U.S. Pat.Nos. 6,346,403, 6,441,274 and 6,664,445 disclose plant amino acidbiosynthetic enzymes. International Patent Appl. Pub. No. WO 98/45458describes synthetic seed proteins having a higher percentage ofessential amino acids than wild-type.

U.S. Pat. No. 5,633,436 discloses plants comprising a higher content ofsulfur-containing amino acids; U.S. Pat. No. 5,885,801 discloses plantscomprising a high threonine content; U.S. Pat. No. 5,885,802 disclosesplants comprising a high methionine content; U.S. Pat. No. 5,912,414discloses plants comprising a high methionine content; U.S. Pat. No.5,990,389 discloses plants comprising a high lysine content; U.S. Pat.No. 6,459,019 discloses plants comprising an increased lysine andthreonine content; International Patent Appl. Pub. No. WO 98/42831discloses plants comprising a high lysine content; International PatentAppl. Pub. No. WO 96/01905 discloses plants comprising a high threoninecontent; and International Patent Appl. Pub. No. WO 95/15392 disclosesplants comprising a high lysine content.

M. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Barren Plants: Plants that are barren, i.e., lack an ear with grain orhave an ear with only a few scattered kernels.

Cg: Colletotrichum graminicola rating. Rating times 10 is approximatelyequal to percent total plant infection.

CLN: Corn Lethal Necrosis (combination of Maize chlorotic mottle virusand Maize dwarf mosaic virus) rating. Numerical ratings are based on aseverity scale where 1=most resistant to 9=susceptible.

Cn: Corynebacterium nebraskense rating. Rating times 10 is approximatelyequal to percent total plant infection.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Cz: Cercospora zeae-maydis rating. Rating times 10 is approximatelyequal to percent total plant infection.

Dgg: Diatraea grandiosella girdling rating (values are percent plantsgirdled and stalk lodged).

Diploid: A cell or organism having two sets of chromosomes.

Dropped Ears: Ears that have fallen from the plant to the ground.

Dsp: Diabrotica species root ratings (1=least affected to 9=severepruning).

Ear-Attitude: The attitude or position of the ear at harvest scored asupright, horizontal, or pendant.

Ear-Cob Color: The color of the cob, scored as white, pink, red, orbrown.

Ear-Cob Diameter: The average diameter of the cob measured at themidpoint.

Ear-Cob Strength: A measure of mechanical strength of the cobs tobreakage, scored as strong or weak.

Ear-Diameter: The average diameter of the ear at its midpoint.

Ear-Dry Husk Color: The color of the husks at harvest scored as buff,red, or purple.

Ear-Fresh Husk Color: The color of the husks 1 to 2 weeks afterpollination scored as green, red, or purple.

Ear-Husk Bract: The length of an average husk leaf scored as short,medium, or long.

Ear-Husk Cover: The average distance from the tip of the ear to the tipof the husks, minimum value no less than zero.

Ear-Husk Opening: An evaluation of husk tightness at harvest scored astight, intermediate, or open.

Ear-Length: The average length of the ear.

Ear-Number Per Stalk: The average number of ears per plant.

Ear-Shank Internodes: The average number of internodes on the ear shank.

Ear-Shank Length: The average length of the ear shank.

Ear-Shelling Percent: The average of the shelled grain weight divided bythe sum of the shelled grain weight and cob weight for a single ear.

Ear-Silk Color: The color of the silk observed 2 to 3 days after silkemergence scored as green-yellow, yellow, pink, red, or purple.

Ear-Taper (Shape): The taper or shape of the ear scored as conical,semi-conical, or cylindrical.

Ear-Weight: The average weight of an ear.

Early Stand: The percent of plants that emerge from the ground asdetermined in the early spring.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

ER: Ear rot rating (values approximate percent ear rotted).

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Final Stand Count: The number of plants just prior to harvest.

GDUs: Growing degree units which are calculated by the Barger Method,where the heat units for a 24-h period are calculated as GDUs=[(Maximumdaily temperature+Minimum daily temperature)/2]-50. The highest maximumdaily temperature used is 86° F. and the lowest minimum temperature usedis 50° F.

GDUs to Shed: The number of growing degree units (GDUs) or heat unitsrequired for a variety to have approximately 50% of the plants sheddingpollen as measured from time of planting. GDUs to shed is determined bysumming the individual GDU daily values from planting date to the dateof 50% pollen shed.

GDUs to Silk: The number of growing degree units for a variety to haveapproximately 50% of the plants with silk emergence as measured fromtime of planting. GDUs to silk is determined by summing the individualGDU daily values from planting date to the date of 50% silking.

Genetic Complement: An aggregate of nucleotide sequences, the expressionof which sequences defines the phenotype in corn plants, or componentsof plants including cells or tissue.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Hct: Helminthosporium carbonum race 2 rating. Rating times 10 isapproximately equal to percent total plant infection.

Hc3: Helminthosporium carbonum race 3 rating. Rating times 10 isapproximately equal to percent total plant infection.

Hm: Helminthosporium maydis race 0 rating. Rating times 10 isapproximately equal to percent total plant infection.

Ht1: Helminthosporium turcicum race 1 rating. Rating times 10 isapproximately equal to percent total plant infection.

Ht2: Helminthosporium turcicum race 2 rating. Rating times 10 isapproximately equal to percent total plant infection.

HtG: Chlorotic-lesion type resistance. “+” indicates the presence of Htchlorotic-lesion type resistance; “−” indicates absence of Htchlorotic-lesion type resistance; and “+/−” indicates segregation of Htchlorotic-lesion type resistance. Rating times 10 is approximately equalto percent total plant infection.

Kernel-Aleurone Color: The color of the aleurone scored as white, pink,tan, brown, bronze, red, purple, pale purple, colorless, or variegated.

Kernel-Cap Color: The color of the kernel cap observed at dry stage,scored as white, lemon-yellow, yellow, or orange.

Kernel-Endosperm Color: The color of the endosperm scored as white, paleyellow, or yellow.

Kernel-Endosperm Type: The type of endosperm scored as normal, waxy, oropaque.

Kernel-Grade: The percent of kernels that are classified as rounds.

Kernel-Length: The average distance from the cap of the kernel to thepedicel.

Kernel-Number Per Row: The average number of kernels in a single row.

Kernel-Pericarp Color: The color of the pericarp scored as colorless,red-white crown, tan, bronze, brown, light red, cherry red, orvariegated.

Kernel-Row Direction: The direction of the kernel rows on the ear scoredas straight, slightly curved, spiral, or indistinct (scattered).

Kernel-Row Number: The average number of rows of kernels on a singleear.

Kernel-Side Color: The color of the kernel side observed at the drystage, scored as white, pale yellow, yellow, orange, red, or brown.

Kernel-Thickness: The distance across the narrow side of the kernel.

Kernel-Type: The type of kernel scored as dent, flint, or intermediate.

Kernel-Weight: The average weight of a predetermined number of kernels.

Kernel-Width: The distance across the flat side of the kernel.

Kz: Kabatiella zeae rating. Rating times 10 is approximately equal topercent total plant infection.

Leaf-Angle: Angle of the upper leaves to the stalk scored as upright (0to 30 degrees), intermediate (30 to 60 degrees), or lax (60 to 90degrees).

Leaf-Color: The color of the leaves 1 to 2 weeks after pollinationscored as light green, medium green, dark green, or very dark green.

Leaf-Length: The average length of the primary ear leaf.

Leaf-Longitudinal Creases: A rating of the number of longitudinalcreases on the leaf surface 1 to 2 weeks after pollination. Creases arescored as absent, few, or many.

Leaf-Marginal Waves: A rating of the waviness of the leaf margin 1 to 2weeks after pollination, rated as none, few, or many.

Leaf-Number: The average number of leaves of a mature plant. Countingbegins with the cotyledonary leaf and ends with the flag leaf.

Leaf-Sheath Anthocyanin: A rating of the level of anthocyanin in theleaf sheath 1 to 2 weeks after pollination, scored as absent,basal-weak, basal-strong, weak, or strong.

Leaf-Sheath Pubescence: A rating of the pubescence of the leaf sheath.Ratings are taken 1 to 2 weeks after pollination and scored as light,medium, or heavy.

Leaf-Width: The average width of the primary ear leaf measured at itswidest point.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

LSS: Late season standability (values times 10 approximate percentplants lodged in disease evaluation plots).

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Moisture: The moisture of the grain at harvest.

On1: Ostrinia nubilalis 1st brood rating (1=resistant to 9=susceptible).

On2: Ostrinia nubilalis 2nd brood rating (1=resistant to 9=susceptible).

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Regeneration: The development of a plant from tissue culture.

Relative Maturity: A maturity rating based on regression analysis. Theregression analysis is developed by utilizing check hybrids and theirpreviously established day rating versus actual harvest moistures.Harvest moisture on the hybrid in question is determined and thatmoisture value is inserted into the regression equation to yield arelative maturity.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Root Lodging: Root lodging is the percentage of plants that root lodge.A plant is counted as root lodged if a portion of the plant leans fromthe vertical axis by approximately 30 degrees or more.

Royal Horticultural Society (RHS) Color Chart Value: The RHS Color Chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightness,and saturation. A color is precisely named by the RHS Color Chart byidentifying the group name, sheet number, and letter, e.g.,Yellow-Orange Group 19A or Red Group 41B.

Seedling Color: Color of leaves at the 6 to 8 leaf stage.

Seedling Height: Plant height at the 6 to 8 leaf stage.

Seedling Vigor: A visual rating of the amount of vegetative growth on a1 to 9 scale, where 1 equals best. The score is taken when the averageentry in a trial is at the fifth leaf stage.

Selection Index: The selection index gives a single measure of hybrid'sworth based on information from multiple traits. One of the traits thatis almost always included is yield. Traits may be weighted according tothe level of importance assigned to them.

Self-Pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing or genetic engineering ofa locus, wherein essentially all of the morphological and physiologicalcharacteristics of a corn variety are recovered in addition to thecharacteristics of the single locus.

Sr: Sphacelotheca reiliana rating is actual percent infection.

Stalk-Anthocyanin: A rating of the amount of anthocyanin pigmentation inthe stalk. The stalk is rated 1 to 2 weeks after pollination as absent,basal-weak, basal-strong, weak, or strong.

Stalk-Brace Root Color: The color of the brace roots observed 1 to 2weeks after pollination as green, red, or purple.

Stalk-Diameter: The average diameter of the lowest visible internode ofthe stalk.

Stalk-Ear Height: The average height of the ear measured from the groundto the point of attachment of the ear shank of the top developed ear tothe stalk.

Stalk-Internode Direction: The direction of the stalk internode observedafter pollination as straight or zigzag.

Stalk-Internode Length: The average length of the internode above theprimary ear.

Stalk Lodging: The percentage of plants that did stalk lodge. Plants arecounted as stalk lodged if the plant is broken over or off below theear.

Stalk-Nodes With Brace Roots: The average number of nodes having braceroots per plant.

Stalk-Plant Height: The average height of the plant as measured from thesoil to the tip of the tassel.

Stalk-Tillers: The percent of plants that have tillers. A tiller isdefined as a secondary shoot that has developed as a tassel capable ofshedding pollen.

Staygreen: Staygreen is a measure of general plant health near the timeof black layer formation (physiological maturity). It is usuallyrecorded at the time the ear husks of most entries within a trial haveturned a mature color. Scoring is on a 1 to 9 basis where 1 equals best.

STR: Stalk rot rating (values represent severity rating of 1=25% ofinoculated internode rotted to 9=entire stalk rotted and collapsed).

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

SVC: Southeastern Virus Complex (combination of Maize chlorotic dwarfvirus and Maize dwarf mosaic virus) rating; numerical ratings are basedon a severity scale where 1=most resistant to 9=susceptible.

Tassel-Anther Color: The color of the anthers at 50% pollen shed scoredas green-yellow, yellow, pink, red, or purple.

Tassel-Attitude: The attitude of the tassel after pollination scored asopen or compact.

Tassel-Branch Angle: The angle of an average tassel branch to the mainstem of the tassel scored as upright (less than 30 degrees),intermediate (30 to 45 degrees), or lax (greater than 45 degrees).

Tassel-Branch Number: The average number of primary tassel branches.

Tassel-Glume Band: The closed anthocyanin band at the base of the glumescored as present or absent.

Tassel-Glume Color: The color of the glumes at 50% shed scored as green,red, or purple.

Tassel-Length: The length of the tassel measured from the base of thebottom tassel branch to the tassel tip.

Tassel-Peduncle Length: The average length of the tassel peduncle,measured from the base of the flag leaf to the base of the bottom tasselbranch.

Tassel-Pollen Shed: A visual rating of pollen shed determined by tappingthe tassel and observing the pollen flow of approximately five plantsper entry. Rated on a 1 to 9 scale where 9=sterile, 1=most pollen.

Tassel-Spike Length: The length of the spike measured from the base ofthe top tassel branch to the tassel tip.

Test Weight: Weight of the grain in pounds for a given volume (bushel)adjusted to 15.5% moisture.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a corn plant by transformation orsite-specific modification.

Yield: Yield of grain at harvest adjusted to 15.5% moisture.

N. Deposit Information

A deposit of corn line SHY-6RSNL196, disclosed above and recited in theclaims, has been made with the American Type Culture Collection (ATCC),10801 University Blvd., Manassas, Va. 20110-2209. The date of depositfor corn line SHY-6RSNL196 was Feb. 19, 2020. The accession numbers forthose deposited seeds of corn line SHY-6RSNL196 is ATCC Accession NumberPTA-126668. Upon issuance of a patent, all restrictions upon the depositwill be removed, and the deposit is intended to meet all of therequirements of 37 C.F.R. §§ 1.801-1.809. The deposit will be maintainedin the depository for a period of 30 years, or 5 years after the lastrequest, or for the effective life of the patent, whichever is longer,and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed:
 1. A corn plant comprising at least a first set of the chromosomes of corn line SHY-6RSNL196, a sample of seed of said line having been deposited under ATCC Accession Number PTA-126668.
 2. A corn seed that produces the plant of claim
 1. 3. The plant of claim 1, wherein the plant is a plant of said corn line SHY-6RSNL196.
 4. The plant of claim 1, wherein the plant is a hybrid corn plant.
 5. The seed of claim 2, wherein the seed is a seed of said corn line SHY-6RSNL196.
 6. The seed of claim 2, wherein the seed is a seed of a hybrid corn plant.
 7. A plant part of the plant of claim 1, wherein the plant part comprises a cell of said plant.
 8. A corn plant having all the physiological and morphological characteristics of the plant of claim
 3. 9. A tissue culture of regenerable cells of the plant of claim
 1. 10. A method of vegetatively propagating the corn plant of claim 1, the method comprising the steps of: (a) collecting tissue capable of being propagated from the plant of claim 1; and (b) propagating a corn plant from said tissue.
 11. A method of introducing a trait into a corn line, the method comprising: (a) utilizing as a recurrent parent the plant of claim 3 by crossing said plant with a donor corn plant that comprises a trait to produce F₁ progeny; (b) selecting an F₁ progeny that comprises the trait; (c) backcrossing the selected F₁ progeny with a plant of the same corn line used as the recurrent parent in step (a) to produce a backcross progeny; (d) selecting a backcross progeny comprising the trait and otherwise comprising the morphological and physiological characteristics of the recurrent parent corn line used in step (a); and (e) repeating steps (c) and (d) three or more times to produce a selected fourth or higher backcross progeny.
 12. A corn plant produced by the method of claim 11, wherein said plant comprises the trait and otherwise comprises all of the morphological and physiological characteristics of corn line SHY-6RSNL196.
 13. A method of producing a corn plant comprising an added trait, the method comprising introducing a transgene conferring the trait into the plant of claim
 1. 14. A corn plant produced by the method of claim
 13. 15. A corn plant comprising at least a first set of the chromosomes of corn line SHY-6RSNL196, a sample of seed of said line having been deposited under ATCC Accession Number PTA-126668, further comprising a transgene.
 16. The plant of claim 15, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism, and modified protein metabolism.
 17. A corn plant comprising at least a first set of the chromosomes of corn line SHY-6RSNL196, a sample of seed of said line having been deposited under ATCC Accession Number PTA-126668, further comprising a single locus conversion.
 18. The plant of claim 17, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism, and modified protein metabolism.
 19. A method for producing a seed of a corn plant derived from corn line SHY-6RSNL196, the method comprising the steps of: (a) crossing the corn plant of claim 1 with itself or a second corn plant; and (b) allowing a seed of a line SHY-6RSNL196-derived corn plant to form.
 20. A method of producing a seed of a line SHY-6RSNL196-derived corn plant, the method comprising the steps of: (a) producing a line SHY-6RSNL196-derived corn plant from a seed produced by crossing the corn plant of claim 1 with itself or a second corn plant; and (b) crossing the line SHY-6RSNL196-derived corn plant with itself or a different corn plant to obtain a seed of a further line SHY-6RSNL196-derived corn plant.
 21. The method of claim 20, the method further comprising repeating said producing and crossing steps of (a) and (b) using the seed from said step (b) for producing the plant according to step (a) for at least one generation to produce a seed of an additional line SHY-6RSNL196-derived corn plant.
 22. A method of producing an ear of corn, the method comprising: (a) obtaining the plant of claim 1, wherein the plant has been cultivated to maturity; and (b) collecting an ear of corn from the plant. 